CN110596072B - Ratio type fluorescent paper chip based on polyester fiber film, preparation method and application - Google Patents
Ratio type fluorescent paper chip based on polyester fiber film, preparation method and application Download PDFInfo
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Abstract
The invention discloses a ratio type fluorescent paper chip based on a polyester fiber membrane, a preparation method and application, belonging to the technical field of material science engineering and microfluidic chips.A CdTe QDs and a RhB are sequentially grafted to the surface of the polyester fiber membrane, and then a surface molecular imprinting technology is adopted to form 2,4-D imprinting holes on the surface of the polyester fiber membrane, so as to prepare fluorescent molecular imprinting paper; installing fluorescent molecularly imprinted paper on a carrier to prepare a ratio type fluorescent paper chip; the invention uses two kinds of fluorescence, compared with a single fluorescence paper chip, the intrinsic self-correction can be realized, so that the detection result is clearer and more accurate; the method has the advantages of simple pretreatment, can be used for detecting the fruits and vegetables after simple cleaning, crushing and filtering, can be used for quickly and auxiliarily detecting 2,4-D in complex fruit and vegetable samples, and has no more limitation on the variety of the detected samples; the detection time is short, the operation is simple and convenient, and the carrying is convenient; the polyester fiber paper is used as the substrate, so that the cost is reduced, and the environment is protected.
Description
Technical Field
The invention belongs to the technical field of material science engineering and microfluidic chips, and particularly relates to a ratio type fluorescent paper chip based on a polyester fiber film, a preparation method and application.
Background
2, 4-Dichlorophenoxyacetic acid (i.e., 2,4-D), is a phenoxyacetic acid-based plant growth regulator that regulates metabolism by stimulating plant growth, and is commonly used in fruits and vegetables. But the protein is difficult to degrade and is very easy to remain in fruits and vegetables, so that the protein may directly or indirectly enter human bodies through food chains to threaten human health, and the protein has important significance in detecting 2,4-D in fruits and vegetables.
The commonly used 2,4-D detection technology is high performance liquid chromatography, but the method is difficult to popularize due to the reasons of high precision and expensive equipment, long detection time, complex pretreatment and the like, and the detection application of the 2,4-D is severely limited, and the 2,4-D is very easy to remain in fruits and vegetables and causes harm to human bodies, so that the establishment of the 2,4-D detection method which is high in efficiency, convenient to carry, low in cost and environment-friendly is very necessary.
Disclosure of Invention
In order to solve the problems, the invention provides a ratio type fluorescent paper chip based on a polyester fiber film, a preparation method and application thereof, wherein the ratio type fluorescent paper chip can effectively detect 2,4-D, is efficient and rapid, is low in cost, is environment-friendly and is convenient to carry.
The first purpose of the invention is to provide a preparation method of ratio type fluorescent paper chip based on polyester fiber film, comprising the following steps:
sequentially grafting CdTe QDs and RhB (rhodamine B) to the surface of the polyester fiber membrane, and forming 2,4-D imprinted holes on the surface of the polyester fiber membrane by adopting a surface molecular imprinting technology to prepare fluorescent Molecularly Imprinted Paper (MIP); and (3) mounting the fluorescent molecularly imprinted paper on a carrier to prepare the ratio type fluorescent paper chip.
Preferably, the preparation method of the ratio type fluorescent paper chip for rapidly detecting 2,4-D is characterized by comprising the following steps:
s1: acidizing the polyester fiber film, immersing the acidized polyester fiber film into a mixed solution of CdTe QDs and EDC, adding NHS solution, standing, oscillating for 15-17 h, and cleaning to obtain the polyester fiber film grafted with CdTe QDs;
s2: soaking the S1 polyester fiber film grafted with CdTe QDs into an ethanol/TEOS (tetraethyl orthosilicate) mixed solution, oscillating for 3-5 h in a dark place, adding RhB to perform oscillation reaction for 20-40 min, sequentially adding 2,4-D, APTES (3-aminopropyltriethoxysilane), ammonia water and a second batch of TEOS, continuing oscillation reaction for 3-5 h, and eluting for 2,4-D by using a methanol and acetic acid mixed solution after the reaction is finished to prepare the fluorescent molecularly imprinted paper;
s3: and (3) mounting the fluorescent molecularly imprinted paper on a paper chip carrier to prepare the ratio type fluorescent paper chip.
Preferably, the polyester fiber membrane acidification treatment in S1 specifically comprises the following steps: and (3) immersing the polyester fiber film into 0.2mol/L HCl, oscillating for 15-25 min, and washing with water to obtain the polyester fiber film subjected to acidification treatment.
Preferably, the mixed solution of CdTe QDs and EDC in S1 means that CdTe QDs and MES buffer solution dissolved with 20mg/mL EDC are mixed, the NHS solution means MES buffer solution dissolved with 10mg/mL EDC, and the ratio of CdTe QDs: EDC/MES: the volume ratio of NHS/MES is 3-5: 2: 2.
preferably, the ratio of ethanol in S2: TEOS: the dosage ratio of RhB is 5-10 mL: 25 μ L of: 3mg, RhB: the mass ratio of 2,4-D is 3: 4-6, and the mass ratio of 2,4-D is: the dosage ratio of APTES is 4-6 mg: 25 μ L, APTES: ammonia water: the second batch TEOS had a volume ratio of 1: 2: 1.
preferably, the step of S2 elution 2,4-D means that the polyester fiber membrane after reaction is firstly washed by water, then is soaked in methanol and acetic acid solution with the concentration of 0.01M, is eluted for 1 hour, and is washed by water, wherein the ratio of methanol: the volume ratio of acetic acid is 4: 1.
preferably, in S3, the conditions for mounting the fluorescent molecularly imprinted paper on the paper chip carrier are as follows: and naturally airing the fluorescent molecular imprinting paper, and mounting the fluorescent molecular imprinting paper on a carrier in a dark condition, wherein the carrier in S3 is a three-layer microfluidic paper chip obtained by folding cellulose paper obtained by processing hydrophobic wax.
Preferably, the polyester fiber film is DL 42.
The second purpose of the invention is to provide the ratio type fluorescent paper chip prepared by the preparation method.
The third purpose of the invention is to provide the application of the ratio type fluorescent paper chip in 2,4-D detection.
Compared with the prior art, the invention has the following beneficial effects:
2,4-D is taken as a template molecule, a surface molecular imprinting technology is applied, so that the template is easy to elute, the selectivity to the 2,4-D is ensured, and polyester fiber paper is taken as a substrate, so that the cost is reduced, and the environment is protected; by using two types of fluorescence, compared with a single fluorescent paper chip, the ratio fluorescence detection can realize internal self-correction, so that the detection result is clearer and more accurate; the method has the advantages of simple pretreatment, can be used for detecting the fruits and vegetables after simple cleaning, crushing and filtering, can be used for quickly and auxiliarily detecting 2,4-D in complex fruit and vegetable samples, and has no more limitation on the variety of the detected samples; in addition, the invention has the advantages of short detection time, high detection speed, simple operation and convenient carrying.
Drawings
FIG. 1 is a flow chart of the preparation of a ratiometric fluorescent paper chip based on a polyester fiber membrane provided by the invention;
wherein A to G are synthetic processes, and the attached figures are chemical structural formulas of related substances: functional monomer APTES (3-aminopropyltriethoxysilane), cross-linking agent TEOS (tetraethoxysilane), (rhodamine B) RhB, template molecule 2, 4-D;
FIG. 2 is the examination result of the binding capacity of different papers and fluorescent materials;
wherein (a) the fluorescence intensity before and after elution of five different papers combined with CdTe QDs; (b) the fluorescence intensity before and after elution of the five papers combined with RhB; (c) fluorescence intensity before and after elution of three types of polyester fiber film paper (including DL42) combined with CdTe QDs; (d) fluorescence intensities before and after RhB elution were combined with three polyester fiber film papers (including DL 42);
FIG. 3 is a view of the grafting principle of CdTe QDs (a) and RhB (b) to DL42 paper in example 1;
FIG. 4 is an SEM photograph of the MIP prepared in example 1; wherein, the original bare paper and the CdTe QDs single fluorescent paper chip are used as the contrast;
FIG. 5 shows the response time (a) and the fluorescence titration spectrum (b) of the ratiometric fluorescent paper chip prepared in example 1.
Detailed Description
In order to make the technical solutions of the present invention better understood and implemented by those skilled in the art, the present invention is further described below with reference to the following specific embodiments and the accompanying drawings, but the embodiments are not meant to limit the present invention.
The experimental methods and the detection methods described in the following examples are all conventional methods unless otherwise specified; the reagents and materials are commercially available, unless otherwise specified.
Example 1
A method for preparing a ratio-type fluorescent paper chip based on a polyester fiber film, as shown in fig. 1, comprising the following steps:
s1: grafting of CdTe QDs onto paper
Firstly, preparing 100mL of 1mM MES buffer solution, enabling the pH value to be 5.2, respectively preparing 20mg/mL EDC solution and 10mg/L NHS solution by taking the MES buffer solution as a solvent for later use, respectively preparing 20mL of each 20 mg/L EDC solution, cutting polyester fiber membrane paper with the model of DL42 to be 1cm multiplied by 1cm, acidizing the polyester fiber membrane paper with 20mL of 0.2mol/L HCl, pouring off the HCl after shaking for 20min, washing the polyester fiber membrane paper with ultrapure water to remove residual HCl solution, mixing 15mL of CdTe QDs solution with 6mL of EDC, shaking for 10min, pouring the mixture into a culture dish containing the acidized fiber paper, adding 6mL of NHS, standing for 1h, shaking for 16h, and washing with the ultrapure water for three times to remove redundant quantum dots. The process was protected from light to prevent fluorescence quenching.
S2: preparation of MIPs
After the above treatment, 20mL of ethanol and 50. mu.L of TEOS were added, and the mixture was shaken for 4 hours. 6mg of RhB solid was added and reacted for half an hour with shaking. Then, 10mg of template molecule (2,4-D), 50. mu.L of APTES, 100. mu.L of ammonia water and 50. mu.L of TEOS were added in this order, and the reaction was continued for 4 hours with shaking. After rinsing with ultrapure water three times, the template was eluted with 8mL of methanol and 2mL of 0.01M acetic acid for 1 hour, and then rinsed with water three times to obtain MIP, which was stored at 4 ℃ in the dark.
S3: mounting MIPs onto a paper chip carrier
And (3) folding cellulose paper obtained by processing hydrophobic wax to obtain a three-layer microfluidic paper chip carrier, baking the three-layer microfluidic paper chip carrier for 30s at 50 ℃, mounting the MIP on the carrier, and storing the MIP for later use at 4 ℃ to obtain the ratio type fluorescent paper chip.
Example 2
A preparation method of a ratio type fluorescent paper chip based on a polyester fiber film comprises the following steps:
s1: grafting of CdTe QDs onto paper
Firstly, preparing 100mL of 1mM MES buffer solution, enabling the pH value to be 5.2, respectively preparing 20mg/mL EDC solution and 10mg/L NHS solution by taking the MES buffer solution as a solvent for later use, respectively preparing 20mL of each 20mg/mL EDC solution and 10mg/L NHS solution, cutting polyester fiber membrane paper with the model of DL42 to be 1cm multiplied by 1cm, acidizing the polyester fiber membrane paper with 20mL of 0.2mol/L HCl, pouring out the HCl after shaking for 15min, washing with ultrapure water to remove residual HCl solution, mixing 15mL of CdTe QDs solution and 6mL of EDC, shaking for 10min, pouring the mixture into a culture dish containing the acidized fiber paper, adding 6mL of NHS, standing for 1h, shaking for 15h, and washing with the ultrapure water for three times to remove redundant quantum dots. The process was protected from light to prevent fluorescence quenching.
S2: preparation of MIPs
After the treatment, 15mL of ethanol and 50 muL of TEOS are added, the mixture is oscillated for 3h, 6mg of RhB solid is added for oscillation reaction for 20min, then 12mg of template molecule (2,4-D), 50 muL of APTES, 100 muL of ammonia water and 50 muL of TEOS are sequentially added, oscillation reaction is continued for 3h, after the mixture is washed three times by ultrapure water, 8mL of methanol and 2mL of 0.01M acetic acid are added for eluting the template for 1h, and the mixture is washed three times by water to obtain MIP, and the MIP is placed at 4 ℃ and is kept away from light.
S3: mounting MIPs onto a paper chip carrier
And (3) folding cellulose paper obtained by processing hydrophobic wax to obtain a three-layer microfluidic paper chip carrier, baking the three-layer microfluidic paper chip carrier for 30s at 50 ℃, mounting the MIP on the carrier, and storing the MIP for later use at 4 ℃ to obtain the ratio type fluorescent paper chip.
Example 3
A preparation method of a ratio type fluorescent paper chip based on a polyester fiber film comprises the following steps:
s1: grafting of CdTe QDs onto paper
Firstly, preparing 100mL of 1mM MES buffer solution, enabling the pH value to be 5.2, respectively preparing 20mg/mL EDC solution and 10mg/L NHS solution by taking the MES buffer solution as a solvent for later use, respectively preparing 20mL of each 20mg/mL EDC solution and 10mg/L NHS solution, cutting polyester fiber membrane paper with the model of DL42 to be 1cm multiplied by 1cm, acidizing the polyester fiber membrane paper with 20mL of 0.2mol/L HCl, pouring out the HCl after shaking for 25min, washing with ultrapure water to remove residual HCl solution, mixing 9mL of CdTe QDs solution and 6mL of EDC, shaking for 10min, pouring the mixture into a culture dish containing the acidized fiber paper, adding 6mL of NHS, standing for 1h, shaking for 17h, and washing with the ultrapure water for three times to remove redundant quantum dots. The process was protected from light to prevent fluorescence quenching.
S2: preparation of MIPs
After the treatment, 10mL of ethanol and 50 muL of TEOS are added, the mixture is oscillated for 5h, 6mg of RhB solid is added for oscillation reaction for 40min, then 8mg of template molecule (2,4-D), 50 muL of APTES, 100 muL of ammonia water and 50 muL of TEOS are sequentially added, oscillation reaction is continued for 5h, after the mixture is washed three times by ultrapure water, 8mL of methanol and 2mL of 0.01M acetic acid are added for eluting the template for 1h, and the mixture is washed three times by water to obtain MIP, and the MIP is placed at 4 ℃ and is kept away from light.
S3: mounting MIPs onto a paper chip carrier
And (3) folding cellulose paper obtained by processing hydrophobic wax to obtain a three-layer microfluidic paper chip carrier, baking the three-layer microfluidic paper chip carrier for 30s at 50 ℃, mounting the MIP on the carrier, and storing the MIP for later use at 4 ℃ to obtain the ratio type fluorescent paper chip.
The performance of the ratiometric fluorescent paper chips prepared in the above examples 1 to 3 is similar, and the invention will be further described below by taking example 1 as an example.
The binding capacity of different papers and fluorescent materials is examined, as shown in fig. 2, wherein (a) the fluorescence intensity before and after the elution of five different papers bound with CdTe QDs; (b) the fluorescence intensity before and after elution of the five papers combined with RhB; (c) fluorescence intensity before and after elution of three types of polyester fiber film paper (including DL42) combined with CdTe QDs; (d) the fluorescence intensity before and after RhB binding of the three polyester fiber film papers (including DL42) was observed, from the retention degree of fluorescence intensity before and after elution, fig. 2(a) and (b) show that the polyester fiber film paper (DL42) has stronger binding capacity with CdTe QDs and is more suitable for RhB binding than other types of papers, fig. 2(c) and (d) are further investigation on the three polyester fiber film papers (DL42, VL98 and VL78), and research on VL98 and VL78 with front-back difference shows that DL42 has better binding effect with two fluorescent substances, VL98 has front-back difference with CdTe QDs although having better binding effect, and VL78 has no front-back difference but has poor binding degree, and from the above results, DL42 is a suitable base material.
The grafting principle of the CdTe QDs and RhB of the invention and DL42 paper is examined by using urea, SDS (sodium dodecyl sulfate) and NaCl, wherein, FIG. 3(a) is the grafting principle of the CdTe QDs (a) and DL42 paper, FIG. 3(b) is the grafting principle of the RhB and DL42 paper, the urea, SDS and NaCl respectively correspond to hydrogen bond, hydrophobic interaction and electrostatic interaction, the interaction force between the CdTe QDs and DL42 paper obtained from the solute in FIG. 3(a) is mainly electrostatic interaction, while the solute RhB obtained from FIG. 3(b) is mainly combined with paper not only through electrostatic interaction, but also through hydrogen bond and hydrophobic interaction, but mainly electrostatic interaction is adopted.
FIG. 4 is SEM results of original bare paper, CdTe QDs single fluorescent paper chip and MIP, in which the existence of obvious silicon material can be seen, which indicates the successful synthesis of fluorescent molecularly imprinted paper; the result of the response time measurement is shown in fig. 5(a), and the graph shows that the response time is short, and the excellent performance of the rapid detection is reflected; as the concentration of 2,4-D increases, the fluorescence titration spectrum is shown in FIG. 5(b), and the change of two fluorescence signals indicates that the ratio type fluorescent paper chip prepared by the invention can be used for detecting 2,4-D and has better sensitivity.
It will be apparent to those skilled in the art that various changes and modifications may be made in the present invention without departing from the spirit and scope of the invention. Thus, it is intended that such changes and modifications be included within the scope of the appended claims and their equivalents.
Claims (10)
1. A preparation method of a ratio type fluorescent paper chip based on a polyester fiber film is characterized by comprising the following steps:
sequentially grafting CdTe QDs and RhB onto the surface of the polyester fiber membrane, and forming 2,4-D imprinted holes on the surface of the polyester fiber membrane by adopting a surface molecular imprinting technology to prepare fluorescent molecularly imprinted paper; and (3) mounting the fluorescent molecularly imprinted paper on a carrier to prepare the ratio type fluorescent paper chip.
2. The method for preparing the ratio type fluorescent paper chip based on the polyester fiber film as claimed in claim 1, which is characterized by comprising the following steps:
s1: acidizing the polyester fiber film, immersing the acidized polyester fiber film into a mixed solution of CdTe QDs and EDC, adding NHS solution, standing, oscillating for 15-17 h, and cleaning to obtain the polyester fiber film grafted with CdTe QDs;
s2: soaking the S1 polyester fiber membrane grafted with CdTe QDs in an ethanol/TEOS mixed solution, oscillating for 3-5 h in a dark place, adding RhB to perform oscillation reaction for 20-40 min, sequentially adding 2,4-D, APTES, ammonia water and a second batch of TEOS, continuing to perform oscillation reaction for 3-5 h, and eluting 2,4-D by using a methanol and acetic acid mixed solution after the reaction is finished to prepare the fluorescent molecularly imprinted paper;
s3: and (3) mounting the fluorescent molecularly imprinted paper on a paper chip carrier to prepare the ratio type fluorescent paper chip.
3. The method for preparing the ratio type fluorescent paper chip based on the polyester fiber film as claimed in claim 2, wherein the specific steps of the polyester fiber film acidification treatment in the step S1 are as follows: and (3) immersing the polyester fiber film into 0.2mol/L HCl, oscillating for 15-25 min, and washing with water to obtain the polyester fiber film subjected to acidification treatment.
4. The method for preparing a ratio-type fluorescent paper chip based on polyester fiber membrane as claimed in claim 2, wherein the mixed solution of CdTe QDs and EDC in S1 is prepared by mixing CdTe QDs with MES buffer solution dissolved with 20mg/mL EDC, NHS solution is prepared by mixing MES buffer solution dissolved with 10mg/mL EDC, and the ratio of CdTe QDs: EDC/MES: the volume ratio of NHS/MES is 3-5: 2: 2.
5. the method for preparing the polyester fiber film-based ratiometric fluorescent paper chip of claim 2, wherein the molar ratio of ethanol in S2 is as follows: TEOS: the dosage ratio of RhB is 5-10 mL: 25 μ L of: 3mg, RhB: the mass ratio of 2,4-D is 3: 4-6, and the mass ratio of 2,4-D is: the dosage ratio of APTES is 4-6 mg: 25 μ L, APTES: ammonia water: the second batch TEOS had a volume ratio of 1: 2: 1.
6. the method for preparing the polyester fiber film-based ratio-type fluorescent paper chip as claimed in claim 2, wherein the step of S2 elution 2,4-D means that the polyester fiber film after reaction is firstly washed by water, then is immersed in methanol and acetic acid solution with concentration of 0.01M, is eluted for 1h, and is then washed by water, wherein the ratio of methanol: the volume ratio of acetic acid is 4: 1.
7. the method for preparing the polyester fiber film-based ratiometric fluorescent paper chip of claim 2, wherein the fluorescent molecularly imprinted paper is mounted on the paper chip carrier in S3 under the following conditions: and naturally airing the fluorescent molecular imprinting paper, and mounting the fluorescent molecular imprinting paper on a carrier in a dark condition, wherein the carrier in S3 is a three-layer microfluidic paper chip obtained by folding cellulose paper obtained by processing hydrophobic wax.
8. The method for preparing the polyester fiber film-based ratiometric fluorescent paper chip as claimed in claim 1, wherein the polyester fiber film is of type DL 42.
9. A ratiometric fluorescent paper chip prepared according to any one of the preparation methods of claims 1 to 8.
10. The use of the ratiometric fluorescent paper chip of claim 9 for 2,4-D detection.
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